How to amplify 1kb PCR product

Dr. Duncan Clark duncan at
Mon Mar 2 12:38:14 EST 1998

In article <34FAD3B3.4B82 at>, gzzhjr <gzzhjr at PUBLI
>Hello everyone,
>The length of DNA fragment I want to amplified by PCR is about 1kb,but I
>have no experience with this,could someone tell me the appropriate
>temperature and time in denature,annealing and expansion process?
>Thanks for any information.

For Taq polymerase use the usual extension time allowed is 1kb per
minute used at 72C. Denaturation on a block machine is usually 94C for
30 sec, annealing is for 30 secs at a temperature decided by your oligo
design. Use 2.5u Taq per 100ul PCR. 

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288

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